Purpose: :
The use of immortalized HCEC currently is widespread among celldeath and gene transfer vision research investigators. Thisstudy evaluates characteristics of primary versus immortalizedHCEC in early apoptosis and the response of these cells to genetransfer with empty vector (IZsGW, GFP) and the anti-apoptoticgene Bcl-xL.

Results: :
Early Apoptosis rates peaked in iHCEC 6h and 30h after inductionof apoptosis (up to 50% Ann+PI-), in pHCEC after 60h (up to60% Ann+PI-). There was no significant dose dependency. In addition,pHCEC are much more sensitive to pHAGE-CMV-MCS-IZsGW than iHCEC,showing an IZsGreen expression rate of almost 100% already with0.9x10^5 IU per 48well (iHCEC: max. 50% expression rate with2.25x10^5 IU). The expression rates of AAV2-GFP reached a plateauof 80% in pHCEC 17 days after infection, in iHCEC after 23 daysshowing only 30% expression of GFP. Studying protection of HCECwith Lenti-IZsGr-BclxL resulted in early apoptosis in pHCECof 63% (EV) being significantly reduced to 24% (see figures1 and 2). In iHCEC there was no effect of EV versus BclxL resultingin 15-20% early apoptosis in each.